LOCUS       sequence-13066-e        6398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  AUTHORS   Hayer A, Stoeber M, Ritz D, Engel S, Meyer HH, Helenius A
  TITLE     Caveolin-1 is ubiquitinated and targeted to intralumenal vesicles in
            endolysosomes for degradation.
  JOURNAL   J Cell Biol. 2010 Nov 1. 191(3):615-29.
  PUBMED    21041450
REFERENCE   2  (bases 1 to 6398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2010 Nov 1. 191(3):615-29."
COMMENT     Sequence Label: CAV1-mEGFP/FRT/TO
FEATURES             Location/Qualifiers
     source          1..6398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        203..582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        583..786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     737..757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    788..806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    809..827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     837..861
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1547..1556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1553..1555,1556..1558,1559..2272)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1598..1619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1859..1878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2318..2335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2324..2548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2681..2700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2832..2879
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2887..3909
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2887..3909
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4039..4160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4076..4095)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4130..4149
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4209..4225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4209..4225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4222..4244)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4233..4249
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4257..4263,4264..4281,4282..4287))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4302..4323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4440..4457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4611..5199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4691..4710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5370..6161,6162..6230))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5993..6012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6231..6335)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
       61 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      121 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      181 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      241 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      421 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      481 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      541 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      601 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      661 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      721 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
      841 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      901 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tcgaattcgg
      961 cacgagcttc acccagccac cggccagcat gtctgggggc aaatacgtag actccgaggg
     1021 gcacctctac accgttccca tccgggagca gggcaacatc tacaagccca acaacaaggc
     1081 catggcggag gagatgagcg agaagcaggt gtacgacgcg cacaccaagg aaatcgacct
     1141 ggtcaaccgc gaccccaagc atctcaacga cgacgtggtc aagattgatt ttgaagatgt
     1201 gattgcagaa ccagaaggaa cacacagttt tgatggcatc tggaaggcca gcttcaccac
     1261 cttcactgtg acaaaatact ggttttaccg cttgctgtct gccctctttg gcatcccaat
     1321 ggcactcata tggggcattt actttgccat tctttctttc ctgcacatct gggcagttgt
     1381 gccgtgcatt aagagtttcc tgattgagat tcagtgcatc agccgtgtct attccatcta
     1441 cgtccacacc ttctgtgacc cgttctttga ggctgttggc aaaatattca gcaatatccg
     1501 catcaacatg cagaaagaaa caccgcgggc ccgggatcca ccggtcgcca ccatggtgag
     1561 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1621 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1681 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1741 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1801 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1861 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1921 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1981 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2041 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2101 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2161 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tccttaagga
     2221 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     2281 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2341 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2401 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2461 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2521 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2581 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2641 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2701 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2761 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2821 tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
     2881 ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     2941 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     3001 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     3061 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     3121 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     3181 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     3241 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     3301 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     3361 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     3421 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     3481 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     3541 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     3601 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     3661 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     3721 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     3781 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     3841 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     3901 aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3961 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4021 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4081 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4141 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4201 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4261 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4321 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4381 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4801 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5041 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5161 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5281 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5341 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5401 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5461 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5521 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5581 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5641 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5701 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5761 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5821 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5881 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5941 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6001 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6061 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6121 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6181 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6241 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6301 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6361 gacgtcgacg gatcgggaga tctcccgatc ccctatgg
//
